Gonda M A, Benton C V, Massey R J, Schultz A M
Scan Electron Microsc. 1981(Pt 2):45-62.
Hemocyanin (Hcy) from whelks, Busycon canniculatum has been developed as a visual marker for the identification of virus and cell surface antigens by the correlative techniques of fluorescence microscopy, TEM and SEM. A series of hybridomas producing monoclonal antibodies that allow the identification of type-, group-, and class-specific antigenic determinants on the major envelope glycoprotein of mouse mammary tumor virus (MMTV) gp 52 and two group-specific monoclonal antibodies to MMTV gp 36 have been developed. We used these antibodies in the unlabeled antibody Hcy bridge for immunoelectron microscopy (IEM) and found that each gp 52 antigenic determinant was expressed on virus during all stages of morphogenesis and on the infected cell surface while the gp 36 determinants were not detected. The positive labeling of gp 52 by IEM correlated well with immuno-experiments using the 125I Staph protein A plate binding assay (125IPA). The anti-gp 36 monoclonals in contrast, however, gave positive results only in the 125IPA assay. Hybridomas to murine and primate type C virion envelope proteins [gp 70 and p15(E)] have also been developed. None of the monoclonal antibodies to murine type C virus gp 70 or p15(E) gave positive labeling by IEM when tested with Rauscher murine leukemia virus but two were positive by the 125IPA assay. Monoclonal antibodies to the gp 70 of a baboon type C virus, M-7 were positive in IEM, labeling both the cell surface and viral envelope and reacted with virus in the 125IPA assay. Since monoclonal antibodies provide a much better resolution of the molecular structure and antigenic differences of viruses, interpretations of labeling results are thoroughly discussed. Methods for testing the specificity and titer of monoclonals are presented. The unlabeled antibody Hcy bridge technique and recent advances in methodology are reviewed.
海螺Busycon canniculatum中的血蓝蛋白（Hcy）已被开发为一种视觉标记物，可通过荧光显微镜、透射电子显微镜（TEM）和扫描电子显微镜（SEM）等相关技术来鉴定病毒和细胞表面抗原。现已研制出一系列产生单克隆抗体的杂交瘤，这些单克隆抗体能够识别小鼠乳腺肿瘤病毒（MMTV）主要包膜糖蛋白gp 52上的型特异性、组特异性和类特异性抗原决定簇，以及两种针对MMTV gp 36的组特异性单克隆抗体。我们将这些抗体用于未标记抗体Hcy桥免疫电子显微镜（IEM）技术，发现每个gp 52抗原决定簇在形态发生的所有阶段均在病毒上以及感染细胞表面表达，而未检测到gp 36决定簇。IEM对gp 52的阳性标记与使用125I葡萄球菌蛋白A平板结合试验（125IPA）的免疫实验结果高度相关。然而，相比之下，抗gp 36单克隆抗体仅在125IPA试验中给出阳性结果。也已研制出针对鼠类和灵长类C型病毒粒子包膜蛋白[gp 70和p15（E）]的杂交瘤。当用劳氏鼠白血病病毒进行检测时，针对鼠类C型病毒gp 70或p15（E）的单克隆抗体均未通过IEM给出阳性标记，但有两种通过125IPA试验呈阳性。针对狒狒C型病毒M - 7的gp 70的单克隆抗体在IEM中呈阳性，标记了细胞表面和病毒包膜，并且在125IPA试验中与病毒发生反应。由于单克隆抗体能更好地解析病毒的分子结构和抗原差异，因此对标记结果的解释进行了深入讨论。还介绍了检测单克隆抗体特异性和效价的方法。对未标记抗体Hcy桥技术及该方法的最新进展进行了综述。
